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Aim Amphibians require both terrestrial and aquatic environments to complete their life cycles. Thus, they are subject to complex selection pressures stemming from different environments, and these selection pressures are likely to vary geographically with variation in temperature and precipitation. Studies of genetic differentiation along geographical clines allow identification of footprints of these selection pressures. Location China. Methods To identify possible signatures of local adaptation to particular environmental conditions, we conducted a genome-scan with 20,572 single-nucleotide polymorphisms (SNPs) obtained with restriction site-associated DNA sequencing of pooled samples from 10 Microhyla fissipes populations spanning a 1,398 km long latitudinal gradient in China. Results The results revealed significant genetic differentiation among populations (F-ST =0.090). Numerous outlier SNPs associated with variation in both annual average temperature (n = 69) and precipitation (n = 248) were detected. Main conclusions The degree and pattern of population differentiation in the SNPs supported the hypothesis that these SNPs have been subject to directional natural selection associated with temperature and precipitation and, hence, are reflective of geographically varying local adaptation. Hence, conservation and management plans for Microhyla fissipes should take into account this heterogeneity in genetic constitution in its populations.

In this issue of Molecular Ecology, Tepolt et al. (2021) illustrate how the genetic architecture of adaptation and life history influence invasive success. A marvel of many invasive species is that they are incredibly successful despite evolutionary expectations that they will have low adaptive potential and suffer inbreeding depression due to initially small founding population sizes. Determining the combinations of ecoevolutionary factors that permit this apparent "genetic paradox of invasions" is an ongoing endeavour of invasive species research. Tepolt et al. (2021) study the European green crab in its invasive range on the North American west coast. Following a single introduction into California, this crab quickly spread across a wide latitude gradient, despite low diversity in the original founding population. Adaptation of this crab to clinal variation in temperature appeared largely driven by an inferred chromosomal inversion. This inversion exists as a balanced polymorphism in the European home range of green crabs and is associated with thermal tolerance. Tepolt et al. (2021) therefore demonstrate that adaptive evolution post introduction need not be impeded by bottlenecks if variation at key parts of the genome is available and can be maintained in introduced populations. Moreover, Tepolt et al. (2021) show how chromosomal inversions acting as large-effect loci might facilitate rapid responses to selection in introduced populations.

The predominant phylogenetic patterns within a genome do not always reflect correctly the history of evolutionary divergence and speciation, and the true phylogenetic signal tends to be concentrated within low recombination regions of the genome. In this issue of Molecular Ecology, Hennelly et al. (2021) show that this is also the case for intra-specific relationships that are characterized by considerable gene flow between lineages. The study reconstructs the phylogenetic relationships of Indian and Tibetan wolves with other grey wolf (Canis lupus) populations worldwide, and demonstrates that these two populations represent phylogenetically distinct lineages. This inference was supported by using low-recombination regions of autosomal chromosomes and the X chromosome, which proved to be essential for the correct inference of the lineage splitting order. This study illustrates the power of analytical approaches that implement knowledge of genome evolution patterns to reconstruct complex intra-specific evolutionary relationships. The study is also a compelling example of the application of modern phylogenomic approaches in the identification of evolutionarily significant units for the purpose of species conservation.

Immune defence is a key component of fitness, and individuals are expected to have evolved preferences for mates that ensure immunocompetent offspring. Potential preferences include those for mates with specific heritable immune gene profiles ("good genes") or for immunogenetically dissimilar mates to increase offspring immune gene diversity. The major histocompatibility complex (MHC) is by far the most investigated immune gene in mate choice studies, but we still know very little about its role in sexual selection for genetic benefits. In this issue of Molecular Ecology, Huang et al. capitalize on the extraordinary wealth of behavioural, life history and genetic/genomic data from the free-living Soay sheep population on the Island of Hirta to address this problem. While the authors find evidence of both pre- and postcopulatory MHC-based sexual selection, postcopulatory MHC-dissimilar mate choice is indistinguishable from genome-wide effects, suggesting it is a byproduct of inbreeding avoidance in Soay sheep. The study's comprehensive sampling ensures that inferences are generalizable to the entire population and provides a gold standard for studies investigating immune gene-based sexual selection.

Identifying the molecular mechanisms contributing to phenotypic variation in natural populations is a major goal of molecular ecology. However, the multiple regulatory steps between genotype and phenotype mean that many potential mechanisms can lead to trait divergence. To date, the role of transcriptional regulation in local adaptation has received much focus, as we can readily measure mRNA quantity and have a reasonable grasp of how variation in the expression of many protein-coding genes can influence phenotype. Thus, studying the evolution of protein-coding gene mRNA abundance in candidate tissues has led to successes in detecting the molecular mechanisms underlying local adaptation (reviewed by Hill et al., 2021). However, the contribution of differential splicing of precursor mRNA (pre-mRNA) to adaptive differentiation, as well as the loci controlling this variation, remains largely unexplored in wild populations. In their "From the Cover'" article in this issue of Molecular Ecology, Jacobs and Elmer (2021) reanalyse muscle RNA sequencing (RNA-seq) data to quantify the relative contributions of variation in mRNA quantity (differentially expressed "DE" genes) and splice variant identity (differentially spliced "DS" genes) to parallel divergence of wild "benthic" and "pelagic" ecotypes of a salmonid fish, the Arctic charr (Salvelinus alpinus). They found little overlap in the identity and biological functions of DE and DS genes, suggesting that these two regulatory mechanisms act on different cellular traits to complementarily alter organismal phenotype. Furthermore, many DE and DS genes could be mapped to cis-acting QTL, arguing that some of this regulatory divergence is genetically based. DE and DS genes were also more likely to be "hub genes" than their nondivergent counterparts, hinting that this regulatory variation may have a variety of phenotypic effects. The comparison of three independently evolved pairs of benthic and pelagic charr uncovered greater than expected parallelism in both expression and splicing between ecotypes across different lakes, supporting a role for these molecular phenotypes in adaptive divergence. Overall, the findings of Jacobs and Elmer (2021) highlight the importance of alternative splicing as a potential mechanism underlying local adaptation and provide a framework for others hoping to make the most of their RNA-seq data.

Speciation is often portrayed as an "incomplete" or "incipient" process if two groups of organisms, technically distinguishable either by morphology or genetics, can exchange genes. The ultimate outcome of diversification, given this perspective, is complete reproductive isolation. But an increasing amount of evidence suggests that speciation is rarely complete and inter-fertility between different taxonomically accepted species is consistently maintained. In this issue of Molecular Ecology, Linan et al. (2021) provide results that bridge evolutionary processes from populations to phylogenies that indicate suites of closely related tree species in the Mascarene Islands actively exchange genes, evolving as a nested set of syngameons with a hierarchical pattern of interfertility. The deep insight into diversification provided by this study is particularly powerful because of the genomic scale of the data and the complete taxonomic sampling of an island clade evolving in situ. The prevalence of syngameon dynamics in a broad range of organisms indicates that we should adopt a fluid and comprehensive approach to defining evolutionary units for conservation and research. We should move beyond focusing on single endangered species in evolutionary and ecological isolation from other species but consider the entire network of potentially interfertile species and the potential for future adaptation and innovation, particularly in a human dominated world.

Species with a circannual life cycle need to match the timing of their life history events to the environment to maximize fitness. However, our understanding of how circannual traits such as timing of reproduction are regulated on a molecular level remains limited. Recent studies have implicated that epigenetic mechanisms can be an important part in the processes that regulate circannual traits. Here, we explore the role of DNA methylation in mediating reproductive timing in a seasonally breeding bird species, the great tit (Parus major), using genome‐wide DNA methylation data from individual females that were blood sampled repeatedly throughout the breeding season. We demonstrate rapid and directional changes in DNA methylation within the promoter region of several genes, including a key transcription factor (NR5A1) known from earlier studies to be involved in the initiation of timing of reproduction. Interestingly, the observed changes in DNA methylation at NR5A1 identified here are in line with earlier gene expression studies of reproduction in chicken, indicating that the observed shifts in DNA methylation at this gene can have a regulatory role. Our findings provide an important step towards elucidating the genomic mechanism that mediates seasonal timing of a key life history traits and provide support for the idea that epigenetic mechanisms may play an important role in circannual traits., see also the Perspective by Melanie J. Heckwolf and Britta S. Meyer

DNA metabarcoding is a popular methodology for biodiversity assessment and increasingly used for community level analysis of intraspecific genetic diversity. The evolutionary history of hundreds of specimens can be captured in a single collection vial. However, the method is not without pitfalls, which may inflate or misrepresent recovered diversity metrics. Nuclear pseudogene copies of mitochondrial DNA (numts) have been particularly difficult to control because they can evolve rapidly and appear deceptively similar to true mitochondrial sequences. While the problem of numts has long been recognized for traditional sequencing approaches, the issues they create are particularly evident in metabarcoding in which the identity of individual specimens is generally not known. In this issue of Molecular Ecology Resources, Andujar et al. (2021) provide an easy to implement bioinformatic approach to reduce erroneous sequences due to numts and residual noise in metabarcoding data sets. The metaMATE software designates input sequences as authentic (mtDNA haplotypes) or nonauthentic (numts and erroneous sequences) by comparison to reference data and by analysing nucleotide substitution patterns. Filtering is applied over a range of abundance thresholds and the choice to proceed with a more rigid or less strict sequence removal strategy is at the researchers' discretion. This is a valuable addition to a growing number of complementary tools for improving the reliability of modern biodiversity monitoring.

The technological ability to characterize genetic variation at a genome-wide scale provides an unprecedented opportunity to study the genetic underpinnings and evolutionary mechanisms that promote and sustain biodiversity. The transition from short- to long-read sequencing is particularly promising and allows a more holistic view on any changes in genetic diversity across time and space. Long-read sequencing has tremendous potential but sequencing success strongly depends on the long-range integrity of DNA molecules and therefore on the availability of high-quality tissue samples. With the scope of genomic experiments expanding and wild populations simultaneously disappearing at an unprecedented rate, access to high-quality samples may soon be a major concern for many projects. The need for high-quality biodiversity tissue archives is therefore urgent but sampling and preserving high-quality samples is not a trivial exercise. In this review, I will briefly outline how long-read sequencing can benefit the study of molecular ecology, how this will substantially increase the demand for high-quality tissues and why it is challenging to preserve DNA integrity. I will then provide an overview of preservation approaches and end with a call for support to acknowledge the efforts needed to assemble high-quality tissue archives. In doing so, I hope to simultaneously motivate field biologists to expand sampling practices and molecular biologists to develop (cost) efficient guidelines for the sampling and long-term storage of tissues. A concerted, interdisciplinary, effort is needed to catalogue the genetic variation underlying contemporary biodiversity and will eventually provide a critical resource for future studies.

Historic and modern efforts to understand lichen diversity and evolution have overwhelmingly concentrated on that of the fungal partner, which represents one of the most taxonomically diverse nutritional modes among the Fungi. But what about the algal and cyanobacterial symbionts? An explosion of studies on these cryptic symbionts over the past 20+ years has facilitated a richer understanding of their diversity, patterns of association, and the symbiosis itself. In a From the Cover article in this issue of Molecular Ecology, Dal Forno et al. (2021) provide new insight into one of the most fascinating lichen symbioses. By sequencing cyanobacterial symbionts from over 650 specimens, they reveal the presence of overlooked cyanobacterial diversity, evidence for symbiont sharing among distantly related fungi, and utilize a comparative dating framework to demonstrate temporal discordance among interacting fungal and cyanobacterial lineages.

The archaea that can be readily cultivated in the laboratory are only a small fraction of the total diversity that exists in nature. Although molecular ecology methods, such as metagenomic sequencing, can provide valuable information independent of cell cultivation, it is only through cultivation-based experiments that they may be fully characterized, both for their physiological and ecological properties. Here, we report our efforts towards enriching and isolation of uncultivated archaea from marine sediments using a refined combination of conventional microbial cultivation methods. Initially, cells were retrieved from the sediment samples through a cell extraction procedure and the sediment-free mixed cells were then divided into different size-range fractions by successive filtration through 0.8 µm, 0.6 µm and 0.2 µm membranes. Archaeal 16S rRNA gene analyses indicated noticeable retention of different archaeal groups in different fractions. For each fraction, supplementation with a variety of defined substrates (e.g., methane, sulfate, and lignin) and stepwise dilutions led to highly active enrichment cultures of several archaeal groups with Bathyarchaeota most prominently enriched. Finally, using a roll-bottle technique, three co-cultures consisting of Bathyarchaeota (subgroup-8) and a bacterial species affiliated with either Pseudomonas or Glutamicibacter were obtained. Our results demonstrate that a combination of cell extraction, size fractionation, and roll-bottle isolation methods could be a useful protocol for the successful enrichment and isolation of numerous slow-growing archaeal groups from marine sediments.

Genomic diversity and past population histories are key considerations in the fields of conservation and evolutionary biology. In this issue of Molecular Ecology Resources, Prasad et al. (Mol. Ecol. Resour., 2021) examine how the quality and phylogenetic divergence of reference genomes influences the outcomes of downstream analyses such as diversity and demographic history inference. Using the beluga whale and rowi kiwi as examples (Figure 1), they systematically estimate heterozygosity, runs of homozygosity (ROH), and demographic history (PSMC) using reference genomes of varying quality and phylogenetic divergence from the target species. They show that demographic history analyses are impacted by phylogenetic distance, although this is not pronounced until divergence exceeds 3% from the target species. Similarly, their results imply that heterozygosity estimates are dependent on phylogenetic distance and the method used to perform the estimates, and ROHs are potentially undetectable when a nonconspecific reference is used. This investigation into the role of divergence and quality of reference genomes highlights the impact and potential biases generated by genome selection on downstream analyses, and provides a possible alternative in cross-species scaffolding in instances where a conspecific reference genome is not available.

Sex chromosomes are strange things, and often exhibit unusual patterns of diversity, rates of evolution, and gene regulation (Bachtrog et al., 2011, Mank, 2013). These unique features mean that although sex chromosomes are often a relatively small proportion of the genome, they are best identified and assessed separately from the autosomal majority when carrying out genomic analyses. However, identifying and partitioning genomic regions into sex-linked and autosomal in non-model species can often be quite difficult. In this issue of Molecular Ecology Resources, Nursyifa et al. (2021) provide a useful method that combines sequencing depth information with clustering models to assign sex to samples at the same time as identifying sex-linked scaffolds. This method gives robust results even with more challenging or low-quality data, and thus is particularly promising in studies of non-model organisms.

Arctic and alpine, high latitude and high elevation environments are one of the most stressful environments for species to inhabit. This harshness manifests itself in lower species richness in comparison to more southern vegetation zones (Francis & Currie, 2003). Furthermore, the climatic oscillations-past and predicted-have the most dramatic effect on these ecosystems. For example, in regions of continental ice sheets-the northernmost part of Western Europe and North America-the Arctic species assemblages are no older than a few thousands of years, which is a relatively short period from an evolutionary perspective. Although similar environments may have existed further south during the Ice Age, allowing some preadaptation for the Arctic species, the current habitat is a unique combination of environmental factors such as the climate, soil, bedrock, and photoperiod. Hence, understanding the evolutionary forces shaping Arctic-alpine species will be important for predicting these vulnerable environments' population viability and adaptive potential in the future. In this issue of Molecular Ecology Resources, Nowak et al. (Molecular Ecology Resources) present extensive genome-wide resources for an Arctic-alpine plant Draba nivalis. This adds a valuable new member into the cabbage family models for evolutionary genetics and adaptation studies, to accompany e.g., Arabidopsis (Nature Genetics, 43, 476; Nature, 408, 796), Arabis (Nature Plants, 1, 14023) and Capsella (Nature Genetics, 45, 831). A whole new avenue will open up for molecular ecological studies not only for D. nivalis, but the whole large Draba genus with its diverse ecological and evolutionary characteristics.

The use of high-throughput DNA sequencing (HTS) has revolutionized the assessment of biodiversity in plant and animal communities. There are two main approaches to estimate the identity and the relative species abundance (RSA) in complex mixtures using HTS: amplicon metabarcoding and shotgun metagenomics. While amplicon metabarcoding targets one or a few genomic regions, shotgun metagenomics randomly explores the genome of the species. In this issue of Molecular Ecology Resources, Wagemaker et al. (2021) present a new method, multi-species Genotyping by Sequencing (msGBS), as an alternative middle ground between metabarcoding and metagenomics. They apply the technique to mixtures of plant roots and report the remarkable capacity of msGBS to estimate the RSA. If validated in other laboratories and biological communities, msGBS might become a third method to explore the biodiversity of biological communities, especially of plants, where current techniques are struggling to get sufficient taxonomic resolution.

The quality of genome assemblies has improved rapidly in recent years due to continual advances in sequencing technology, assembly approaches, and quality control. In the field of molecular ecology, this has led to the development of exceptional quality genome assemblies that will be important long-term resources for broader studies into ecological, conservation, evolutionary, and population genomics of naturally occurring species. Moreover, the extent to which a single reference genome represents the diversity within a species varies: pan-genomes will become increasingly important ecological genomics resources, particularly in systems found to have considerable presence-absence variation in their functional content. Here, we highlight advances in technology that have raised the bar for genome assembly and provide guidance on standards to achieve exceptional quality reference genomes. Key recommendations include the following: (a) Genome assemblies should include long-read sequencing except in rare cases where it is effectively impossible to acquire adequately preserved samples needed for high molecular weight DNA standards. (b) At least one scaffolding approach should be included with genome assembly such as Hi-C or optical mapping. (c) Genome assemblies should be carefully evaluated, this may involve utilising short read data for genome polishing, error correction, k-mer analyses, and estimating the percent of reads that map back to an assembly. Finally, a genome assembly is most valuable if all data and methods are made publicly available and the utility of a genome for further studies is verified through examples. While these recommendations are based on current technology, we anticipate that future advances will push the field further and the molecular ecology community should continue to adopt new approaches that attain the highest quality genome assemblies.

Countries' genetic resources and biological variation are extremely valuable. Biodiversity and genetic resources, particularly for endemic species, should be protected. DNA barcoding studies are a useful for identifying unknown taxa and protecting a country's biodiversity. DNA barcodes are small segments of DNA that are taxonomically informative and can be used to for species identification. DNA barcodes have been used to associate life phases, determine cryptic species, and prohibit illegal commerce, among other things. Astragalus argaeus and A. stenosemioides are endemic plant species to Turkey's Mt. Erciyes. To distinguish these two endemic Astagalus, we used two DNA barcodes in the matK region. The current study used A. argaeus, A. stenosemioides, and 23 GenBank sequences of other relative in the genus Astragalus, including one from a closely related species, to infer phylogenetic relationships and DNA barcodes for two endemic plants. The matK gene region could clearly identify A. argaeus and A. stenosemioides from closely related taxa, according to the phylogenetic study. Taxonomic and biodiversity research, as well as molecular ecology and population genetics investigations, will benefit from DNA barcoding surveys to unambiguously differentiate between these two species. Also, by separating the matK DNA gene area, which is one of the molecular features, it is feasible to detect the species, and A. argaeus and A. stenosemioides were successfully barcoded for the first time.

How do you put a name on an unknown piece of DNA? From microbes to mammals, high-throughput metabarcoding studies provide a more objective view of natural communities, overcoming many of the inherent limitations of traditional field surveys and microscopy-based observations (Deiner et al., 2017). Taxonomy assignment is one of the most critical aspects of any metabarcoding study, yet this important bioinformatics task is routinely overlooked. Biodiversity surveys and conservation efforts often depend on formal species inventories: the presence (or absence) of species, and the number of individuals reported across space and time. However, computational workflows applied in eukaryotic metabarcoding studies were originally developed for use with bacterial/archaeal data sets, where microbial researchers rely on one conserved locus (nuclear 16S rRNA) and have access to vast databases with good coverage across most prokaryotic lineages - a situation not mirrored in most multicellular taxa. In this issue of Molecular Ecology Resources, Hleap et al. (2021) carry out an extensive benchmarking exercise focused on taxonomy assignment strategies for eukaryotic metabarcoding studies utilizing the mitochondrial Cytochrome C oxidase I marker gene (COI). They assess the performance and accuracy of software tools representing diverse methodological approaches: from "simple" strategies based on sequence similarity and composition, to model-based phylogenetic and probabilistic classification tools. Contrary to popular assumptions, less complex approaches (BLAST and the QIIME2 feature classifier) consistently outperformed more sophisticated mathematical algorithms and were highly accurate for assigning taxonomy at higher levels (e.g. family). Lower-level assignments at the genus and species level still pose significant challenge for most existing algorithms, and sparse eukaryotic reference databases further limit software performance. This study illuminates current best practices for metabarcoding taxonomy assignments, and underscores the need for community-driven efforts to expand taxonomic and geographic representation in reference DNA barcode databases.

Many forest tree species have characteristics that make the study of their evolutionary ecology complex. For example, they are long-lived and thus have long generation times, and their often large, complex genomes have hampered establishing genomic resources. One way to tackle this challenge is to access multiple complementary data sources and analytical approaches when attempting to infer patterns of adaptive evolution. In the cover article of this issue of Molecular Ecology, Depardieu et al. (2021) combine large amounts of environmental, genomic, dendrochronological, and gene expression data in a common garden to explore the polygenic basis of drought resistance in white spruce (Picea glauca), a long-lived conifer. They identify candidate genes involved in growth and resistance to extreme drought events and show how multiple data sets may deliver complementary evidence to circumvent the manifold challenges of the research field.

Parallel evolution is one of the striking patterns in nature. The presence of repeated evolution of the same phenotypes, suites of traits, and adaptations suggests a strong role for natural selection in shaping biological diversity. The reasoning is straightforward: each instance of repeated evolution makes it less likely that these features evolved neutrally or due to stochastic forces in each population or species. With the growing sequencing capability, we are now poised to examine the genetic basis of parallel evolution in model and nonmodel systems. On pages 4102-4117 of this issue of Molecular Ecology, van Boheemen and Hodgins (2020) provide an exemplar study of this kind, using common ragweed (Ambrosia artemisiifolia; Figure 1a). Their study is noteworthy and ambitious in many respects, and we think will serve as a model for studying parallel adaptation, even in nonmodel species.

Dynamic marine environments can shape complex spatial and temporal patterns in the population connectivity of marine species, and this is often exemplified in species with long larval phases. Here, we used a genotyping-by-sequencing (GBS) approach to examine fine-scale spatial and temporal genomic variation among Dungeness crabCancer magisterlarval recruits sampled in the California Current Ecosystem. Specifically, we compared samples collected during expected- and late-season recruitment time periods within 2 consecutive years (2017 and 2018) at 2 sites in Oregon, USA (Yaquina Bay and Coos Bay). Evidence was found for high gene flow between the expected- and late-season recruits within each year and at both sites based on 1389 neutral loci. In contrast, strong genetic differentiation was observed between these 2 groups within each year and at both sites based on variation at 2 putatively adaptive loci. Contrary to prediction, the magnitude of genetic differentiation between these 2 seasonal groups was greater in 2017 when the Pacific Decadal Oscillation was stronger, upwelling was weaker, and the spring transition was later. Spatial genetic variation was not observed within 2017 or 2018. Comparing across years, expected- and late-season groups were differentiated at putatively adaptive loci. Interestingly, strong genetic differentiation was also observed between late-season groups across years. We found no evidence for cohesive larval dispersal among recruits based on genetic relatedness estimates. Overall, our findings provide evidence for high connectivity within Dungeness crab, but suggest that selective pressures and ocean conditions influence the genetic composition of larval recruits both intra- and inter-annually.

Although cases of independent adaptation to the same dietary niche have been documented in mammalian ecology, the molecular correlates of such shifts are seldom known. Here, we used genomewide analyses of molecular evolution to examine two lineages of bats that, from an insectivorous ancestor, have both independently evolved obligate frugivory: the Old World family Pteropodidae and the neotropical subfamily Stenodermatinae. New genome assemblies from two neotropical fruit bats (Artibeus jamaicensis and Sturnira hondurensis) provide a framework for comparisons with Old World fruit bats. Comparative genomics of 10 bat species encompassing dietary diversity across the phylogeny revealed convergent molecular signatures of frugivory in both multigene family evolution and single-copy genes. Evidence for convergent molecular adaptations associated with frugivorous diets includes the composition of three subfamilies of olfactory receptor genes, losses of three bitter taste receptor genes, losses of two digestive enzyme genes and convergent amino acid substitutions in several metabolic genes. By identifying suites of adaptations associated with the convergent evolution of frugivory, our analyses both reveal the extent of molecular mechanisms under selection in dietary shifts and will facilitate future studies of molecular ecology in mammals.

Chromosomal inversions are increasingly found to differentiate locally adapted populations. This adaptive role is predictable because reduced recombination protects allelic combinations from gene flow. However, we are far from understanding how frequently inversions contribute to local adaptation and how widespread this phenomenon is across species. In a "From the Cover" article in this issue of Molecular Ecology, Huang, Andrew, Owens, Ostevik, and Rieseberg (2020) provide an important step towards this goal not only by finding adaptive inversions in a sunflower ecotype, but also by reversing the approach used to investigate the link between adaptation and inversions. Most studies compare two phenotypes and uncover divergence at a few regions, of which some can subsequently be identified as inversions. In contrast, Huang et al first catalogue putative inversions and then test genotype-environment associations, which allows them to ask systematically whether inversions may be adaptive and in which ecological contexts. They achieve that by revisiting a previous reduced-representation sequencing (RAD-sequencing) data set, demonstrating the suitability of this method to detect inversions in species with limited genomic resources. As such, Huang et al pave the way for a better understanding of the evolutionary role of structural genomic variation and highlight that accounting for inversions in population genomics is now possible, and much needed, in a wider range of organisms.

Understanding the dynamics of communities in space and time requires reconciling ecological and evolutionary processes, including colonization, adaptation, speciation and extinction. In practice, this has been challenging because empirical data obtained by traditional methods and predictive models typically focus on particular processes driving local community assembly and biogeographical structure. In this issue of Molecular Ecology, by using phylogenomics, population genomics and phenomics approaches, Darwell et al. show that ant community assembly on islands is governed by predictable eco-evolutionary trends of geographical range expansion, adaptive radiation and local population decline. The authors provide one of the most robust lines of evidence that the evolutionary progression of island communities may often be directional and repeatable, as predicted by the concept of taxon cycles.

Understanding the process of speciation is a primary goal of evolutionary biology, yet the question of whether speciation can reach completion in the presence of gene flow remains controversial. For more than 50 years, the cichlids of Africa, and more recently those in South and Central America, have served as model systems for the study of speciation in nature. Cichlids are distinguished by their enormous species richness, their diversity of behavioural and trophic adaptations, and their rapid rate of divergence. In both Africa and South and Central America, the repeated interaction of geology, new founder events and adaptive evolution has created a series of natural experiments with speciation occurring both within and between waterbodies of differing ages. In the "From the Cover" paper in this issue of the Journal of Molecular Ecology, Raffini, Schneider, Franchini, Kautt and Meyer move beyond the question of which mechanisms drive speciation, and instead show that divergent morphologies and physiologies translate into adaptive traits. They investigate differences in physiology and gene expression profiles in a benthic/limnetic species pair of Midas cichlidsin a 24,000-year-old Nicaraguan crater lake. While recently diverged, these two species demonstrate significant ecological, but limited genetic differentiation. The authors find that the distinct morphotypes translate into relevant differences in swimming performance and metabolic rates that correspond to differential gene expression profiles. Hence, the authors take an integrative approach examining the impacts of morphological differences on performance and niche partitioning: an approach that can advance our understanding of the drivers of morphological and physiological divergence during speciation.

A major goal in ecology, evolution and conservation biology is understanding how species adapt to changing conditions and using that information to improve conservation actions. Primary to advancing our understanding of adaptation and adaptive potential is determining the causes and consequences of the genetic and phenotypic intraspecific variation that allows for adaptation. However, few studies have been able to link the variation present in molecular process under differing conditions to intraspecific variation in survival. In a From the Cover article in this issue of Molecular Ecology, Harder et al explore the relationship between molecular process and survival to understand the adaptive variation underlying tolerance to low thiamine (vitamin B1, an essential micronutrient) conditions in Atlantic salmon (Salmo salar). Thiamine is required for metabolic functioning, including energy production and nervous and cardiovascular system functioning. By combining controlled breeding and phenotyping with a survival study and transcriptomics, the authors are able to quantify among-family differences in survival under low thiamine conditions. They find wide variation in survival among families, and this survival is linked to differences in gene expression patterns. Their study elucidates the importance of combining different data types to characterize within-population phenotypic variation and understand how that variation may lead to genetic adaptation under stressful conditions.

A major challenge of evolutionary ecology over the next decades is to understand and predict the consequences of the current rapid and important environmental changes on wild populations. Extinction risk of species is linked to populations’ evolutionary potential and to their ability to express adaptive phenotypic plasticity. There is thus a vital need to quantify how selective pressures, quantitative genetics parameters, and phenotypic plasticity, for multiple traits in wild animal populations, may vary with changes in the environment. Here I review our previous research that integrated ecological and evolutionary theories with molecular ecology, quantitative genetics, and long‐term monitoring of individually marked wild animals. Our results showed that assessing evolutionary and plastic changes over time and space, using multi‐trait approaches, under a realistic range of environmental conditions are crucial steps toward improving our understanding of the evolution and adaptation of natural populations. Our current and future work focusses on assessing the limits of adaptive potential by determining the factors constraining the evolvability of plasticity, those generating covariation among genetic variance and selection, as well as indirect genetic effects, which can affect population's capacity to adjust to environmental changes. In doing so, we aim to provide an improved assessment of the spatial and temporal scale of evolutionary processes in wild animal populations.

While metabarcoding of plant DNA from their environment is an exciting method that can supplement inventorying of live plant species, the accuracy and specificity has yet to be fully assessed over complex continuous landscapes. In this work, we evaluate plant community profiles produced via metabarcoding of soil by comparing them to a morphological survey. We assessed plant communities by metabarcoding of soil DNA in 130 sites along ecological gradients (nutrients, succession, moisture) in Denmark using chloroplast trnL region (10–143 bp) primer set and compared the resulting communities to communities produced with a longer nuclear ITS2 region (~216 bp) and a morphological survey. We found that the community variation observed within the morphological survey was well represented by molecular surveys, with significant correlation with both community composition and richness using both primer sets. While the majority of the ITS2 sequences could be assigned to species (over 80%), we had less success with the trnL sequences (70%), which was only possible after restricting the reference database to local species. We conclude that the community profiles produced by metabarcoding can be highly effective in performing large-scale macroecological studies. However, the discovery rates and taxonomic assignments produced via metabarcoding remained inferior to morphological surveys, but manual curation of databases improves the specificity of assignments made by the trnL primers, and improves the accuracy of the assignments made with the ITS2 primers. Finally, we suggest that a greater percentage of named diversity would be recovered by increasing soil sampling with the use of additional universal primer sets.

Understanding how organisms adapt to changing environments is a core focus of research in evolutionary biology. One common mechanism is adaptive introgression, which has received increasing attention as a potential route to rapid adaptation in populations struggling in the face of ecological change, particularly global climate change. However, hybridization can also result in deleterious genetic interactions that may limit the benefits of adaptive introgression. Here, we used a combination of genome-wide quantitative trait locus mapping and differential gene expression analyses between the swordtail fish species Xiphophorus malinche and X. birchmanni to study the consequences of hybridization on thermotolerance. While these two species are adapted to different thermal environments, we document a complicated architecture of thermotolerance in hybrids. We identify a region of the genome that contributes to reduced thermotolerance in individuals heterozygous for X. malinche and X. birchmanni ancestry, as well as widespread misexpression in hybrids of genes that respond to thermal stress in the parental species, particularly in the circadian clock pathway. We also show that a previously mapped hybrid incompatibility between X. malinche and X. birchmanni contributes to reduced thermotolerance in hybrids. Together, our results highlight the challenges of understanding the impact of hybridization on complex ecological traits and its potential impact on adaptive introgression.

We know from human genetic studies that practically all aspects of biology are strongly influenced by the genetic background, as reflected in the advent of “personalized medicine.” Yet, with few exceptions, this is not taken into account when using laboratory populations as animal model systems for research in these fields. Laboratory strains of zebrafish (Danio rerio) are widely used for research in vertebrate developmental biology, behavior, and physiology, for modeling diseases, and for testing pharmaceutic compounds in vivo. However, all of these strains are derived from artificial bottleneck events and therefore are likely to represent only a fraction of the genetic diversity present within the species. Here, we use restriction site-associated DNA sequencing to genetically characterize wild populations of zebrafish from India, Nepal, and Bangladesh, and to compare them to previously published data on four common laboratory strains. We measured nucleotide diversity, heterozygosity, and allele frequency spectra, and find that wild zebrafish are much more diverse than laboratory strains. Further, in wild zebrafish, there is a clear signal of GC-biased gene conversion that is missing in laboratory strains. We also find that zebrafish populations in Nepal and Bangladesh are most distinct from all other strains studied, making them an attractive subject for future studies of zebrafish population genetics and molecular ecology. Finally, isolates of the same strains kept in different laboratories show a pattern of ongoing differentiation into genetically distinct substrains. Together, our findings broaden the basis for future genetic, physiological, pharmaceutic, and evolutionary studies in Danio rerio.

Collecting and storing biological material from wild animals in a way that does not deteriorate data quality for analyses using DNA is instrumental for research in ecology and evolution. Our aims were to collect methods commonly used by researchers for the field collection and long-term storage of blood samples and DNA extracts from wild birds and gather reports on their effectiveness. Personal experiences were collected with an online survey targeted specifically at researchers sampling wild birds. Many researchers experienced problems with blood sample storage but not with DNA extract storage. Storage issues generated problems with obtaining adequate DNA quality and sufficient DNA quantity for the targeted molecular analyses, but were not related to season of blood sampling, access to equipment, transporting samples, temperature and method of blood storage. Final DNA quality and quantity were also not affected by storage time before DNA extraction or the methods used to extract DNA. We discuss practical aspects of field collection and storage and provide some general recommendations, with a list of pros and cons of different preservation methods of avian blood samples and DNA extracts.

A fundamental aspect of evolutionary biology is natural selection on trait variation. Classically, selection has been estimated primarily on external morphological traits such as beak size and coloration, or on easily assayable physiological traits such as heat-tolerance. As technologies and methods improved, evolutionary biologists began examining selection on molecular traits such as protein sequences and cellular processes. In a From the Cover paper in this issue of Molecular Ecology, Ahmad et al. continue this trend by estimating parasite-driven selection on the molecular trait of transcript abundance in a wild population of brown trout (Salmo trutta) by uniquely combining a mark-recapture experimental design with noninvasive RNA sampling. Using transcript abundance to estimate selection allows for many different traits (each unique gene's transcript counts) to be tested in a single experiment, providing the opportunity to examine trends in selection. Ahmad et al. find directional selection strength on transcript counts is generally low and normally distributed. Surprisingly, transcripts under nonlinear selection showed a disruptive selection bias, contradicting previous comparative studies and theoretical work. This highlights the importance of within-generation selection studies, where mechanisms may differ from longer time frames. Their paper also highlights the benefits of a cost-effective 3' RNA sequencing technique to measure gene expression.

Polyploidy, resulting from whole genome duplication, is a widespread phenomenon throughout Eukaryotic kingdoms. It is estimated that 15% of speciation events in plants is due to polyploidization and 16% of plant species encompass ploidy variation. In spite of the evolutionary and economic significance of polyploidy, there is a limited set of tools that would allow routine population genetic and genomic analysis of polyploid systems, in particular for the inference of population diversity and differentiation from large genome-wide data sets (Dufresne et al., 2014; Meirmans, 2020). Such a shortage is striking especially when compared to the rapid development of such tools in diploid systems over the last decade. Consequently, population genomic research in polyploids is still lagging behind diploids, especially in autopolyploids, for example, polyploids with multiple (>2) similar genome copies. In this issue of Molecular Ecology Resources, Shastry et al. (2021) develops the first software specifically designed to infer population structure and ancestry in large genome-wide mixed-ploidy data sets, opening novel opportunities for polyploid population genomic analysis.

Emerging infectious diseases (EIDs) are increasingly recognized as a threat to both biodiversity and human health (Scheele et al., 2019; Wells et al., 2020). But pathogens cannot been seen as unique entities; their intraspecific genetic variability represented in variants, strains, antigenic types or genetic lineages may cause different impacts at the population level (Nelson and Holmes, 2007; Greenspan et al., 2018). The global spread of pathogens has been largely facilitated by globalization of transport, which particularly intensified during the last century (O'Hanlon et al., 2018). As seen with SARS-CoV-2, air travel can rapidly spread a pathogen globally (Wells et al., 2020). Furthermore, after initial introduction subsequent translocations of a pathogen may cause the contact of different variants facilitating the rise of novel genotypes that may have higher pathogenicity or transmissibility (Nelson and Holmes, 2007; Greenspan et al., 2018). Chytridiomycosis is an EID caused by the fungus Batrachochytrium dendrobatidis (Bd), that infects amphibian skin causing population declines to extinction in susceptible species. Now a wildlife pandemic, Bd has been recognized as the single pathogen causing the greatest loss of biodiversity on Earth (Scheele et al., 2019). Recent advances in genetics have made novel tools for pathogen detection and characterization more accessible and reliable (Boyle et al., 2004; Byrne et al., 2019). In this issue of Molecular Ecology Resources, Ghosh et al. (2021) report the development of a new genotyping qPCR assay targeting mitochondrial DNA (mtDNA) of Bd, and based on noninvasive swab samples (Figure 1), discriminate between the two most globally widespread and pathogenic genetic lineages of Bd. Having a better understanding of how the genetic diversity of a pathogen is distributed is crucial to understand their spread patterns and develop timely mitigation strategies.

Leptospirosis is a disease that disproportionately affects impoverished urban communities, but is likely to become more prevalent as changing climate alters flooding regimes. The persistence and transmission of the Leptospira pathogen is reliant on small vertebrate animals, predominantly rodents. In this issue of Molecular Ecology, Peterson et al. demonstrate how changes in rodent diversity and abundances across the complex mosaic of abandonment and recovery investment in post-Katrina New Orleans can predict zoonotic infection prevalence. Understanding the ecological conditions that support persistence and transmission of zoonotic pathogens in urban ecosystems, where they are most likely to affect humans, is critical to effective monitoring and prevention.

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Seventeen polymorphic microsatellite markers were isolated and characterized in Arion vulgaris/lusitanicus, which belongs to the worst European slug pests with serious economic and ecological impact. These markers were tested on 23 individuals collected in a population in Switzerland. Numbers of alleles ranged from 2 to 14 per locus, observed and expected heterozygosities ranged from 0.174 to 0.87, and from 0.162 to 0.903, respectively. These loci were also successfully amplified and were polymorphic in the closely related species A. rufus and A. ater. These loci represent the first highly polymorphic nuclear markers described for A. vulgaris and pave the way for population genetics and molecular ecology research of the important Arion pest slugs.

Logging activities degrade forest habitats across large areas of the tropics, but the impacts on trophic interactions that underpin forest ecosystems are poorly understood. DNA metabarcoding provides an invaluable tool to investigate such interactions, allowing analysis at a far greater scale and resolution than has previously been possible. We analysed the diet of the insectivorous fawn leaf-nosed bat Hipposideros cervinus across a forest disturbance gradient in Borneo, using a dataset of ecological interactions from an unprecedented number of bat-derived faecal samples. Bats predominantly consumed insects from the orders Lepidoptera, Diptera, Blattodea, and Coleoptera, and the taxonomic composition of their diet remained relatively consistent across sites regardless of logging disturbance. There was little difference in the richness of prey consumed per-bat in each logging treatment, indicating potential resilience of this species to habitat degradation. In fact, bats consumed a high richness of prey items, and intensive sampling is needed to reliably compare feeding ecology over multiple sites. Multiple bioinformatic parameters were used, to assess how they altered our perception of sampling completeness. While parameter choice altered estimates of completeness, a very high sampling effort was always required to detect the entire prey community.

DNA metabarcoding is an important tool for molecular ecology. However, its effectiveness hinges on the quality of reference sequence databases and classification parameters employed. Here we evaluate the performance of MiFish 12S taxonomic assignments using a case study of California Current Large Marine Ecosystem fishes to determine best practices for metabarcoding. Specifically, we use a taxonomy cross-validation by identity framework to compare classification performance between a global database comprised of all available sequences and a curated database that only includes sequences of fishes from the California Current Large Marine Ecosystem. We demonstrate that the regional database provides higher assignment accuracy than the comprehensive global database. We also document a tradeoff between accuracy and misclassification across a range of taxonomic cutoff scores, highlighting the importance of parameter selection for taxonomic classification. Furthermore, we compared assignment accuracy with and without the inclusion of additionally generated reference sequences. To this end, we sequenced tissue from 597species using the MiFish 12S primers, adding 252species to GenBank's existing 550 California Current Large Marine Ecosystem fish sequences. We then compared species and reads identified from seawater environmental DNA samples using global databases with and without our generated references, and the regional database. The addition of new references allowed for the identification of 16 additional native taxa representing 17.0% of total reads from eDNA samples, including species with vast ecological and economic value. Together these results demonstrate the importance of comprehensive and curated reference databases for effective metabarcoding and the need for locus-specific validation efforts.

Caiman latirostris has a large geographic distribution, that includes Argentina, Bolivia, Brazil, Paraguay, and Uruguay. In Brazil illegal hunting and land use change have caused population decline, relatively well documented in the last three decades. Due to such circumstances, the estimate of species effective population size might help analyze its viability. Single-sample estimator was used to estimate current effective population size (Ne) of broad-snouted caiman populations in representative areas of the species range in Brazil. For the analyzes, genotypes previously obtained were used for subpopulations of the captive colony of the University of Sao Paulo (USP) and for wild subpopulations. The microsatellites used were Amiμ8, Amiμ11, Amiμ13, Amiμ20, Claμ2, Claμ5, Claμ6, Claμ7, Claμ8, Claμ9 and Claμ10. The 11 loci analyzed produced 18.27 alleles on average. Wild populations showed significant genic and genotypic differentiation among them (p

Increasing studies have begun to focus on biodiversity-productivity relationships for soil microorganisms through molecular ecology methods. However, most of these studies involve controlled experiments, and whether the relationship remains at large spatial scales is still largely unknown. To unravel this issue, archived desert soils from long-term experiments were analysed using high-throughput sequencing, and satellite-derived vegetation datasets were acquired to quantify productivity. Most of the abundant genera were significantly different between low- and high-productivity conditions, and soil bacterial communities were strongly impacted by productivity. Soil bacterial biodiversity, including observed OTUs (operational taxonomic units) and the Chao1, Shannon, and Faith's PD indexes, increased rapidly with productivity at low levels and then reached a relatively stable state, and similar phenomena were observed at multiple taxonomic ranks and for most of the dominant groups. Furthermore, we discovered that the mechanisms resulting in the observed relationship might be ecosystem resource availability in large-scale regions and species competition in local regions. Collectively, these results enhance our understanding of the linkage between belowground microorganisms and aboveground vegetation in arid and semi-arid areas and confirm the potential value of satellite-derived datasets in research on soil microbial diversity at large spatial scales. This article is protected by copyright. All rights reserved.

Pritam Banerjee1, 2, Kathryn A. Stewart3, Caterina M. Antognazza4, Ingrid V. Bunholi5, Kristy Deiner6, Matthew A. Barnes7, Santanu Saha8, Heloise Verdier9, Hideyuki Doi10, Jyoti Prakash Maity2, Michael W.Y. Chan1, Chien Yen Chen2*1Department of Biomedical Sciences, Graduate Institute of Molecular Biology, National Chung Cheng University, 168 University Road, Ming-Shung, Chiayi County 62102, Taiwan2Department of Earth and Environmental Sciences, National Chung Cheng University, 168 University Road, Ming-Shung, Chiayi County 62102, Taiwan.3 Institute of Environmental Science, Leiden University, 2333 CC Leiden, The Netherlands4 Department of Theoretical and Applied Science, University of Insubria, Via J.H. Dunant, 3, 21100, Varese, Italy5 Department of Biology, Indiana State University, Terre Haute, IN 47809, USA6 Department of Environmental Systems Science, ETH Zurich, Universitatstrasse 16, CH-8092 Zurich, Switzerland7 Department of Natural Resources Management, Texas Tech University, Lubbock, TX USA8Post Graduate Department of Botany, Bidhannagar College, Salt Lake City, Kolkata 700064, India9Univ Lyon, Universite Claude Bernard Lyon 1, CNRS, ENTPE, UMR 5023 LEHNA, Villeurbanne, France10Graduate School of Information Science, University of Hyogo, 7-1-28 Minatojima-minamimachi, Chuo-ku, Kobe, 650-0047, JapanAbstract Plant-animal interactions (PAI) represent major channels of energy transfer through ecosystems, where both positive and negative relationships simultaneously contribute to ecosystem functioning. Extinction of a single plant species may have deleterious effects on associated animals and vice-versa, and loss of interactions may occur prior to species-extinction. Monitoring species-interactions is therefore directly related to environmental health and functioning, and studying complex interactions through accurate, cost-effective sampling can aid in the management of detrimental anthropogenic impacts. Conventional PAI monitoring methods (e.g., camera, malaise, and pitfall traps) are potentially invasive, time-consuming, and often unable to achieve species-specific detection. While DNA barcoding of gut contents or bulk samples provides species-specific detection, saves time, and enables simultaneous detection of many taxa, these methods remain potentially invasive and may require the sacrifice of study organisms. Alternatively, non-invasive environmental DNA (eDNA)-based monitoring provides accessible collection of biological signatures from the environment (air, water, soil) that can elucidate PAI. Environmental DNA methods have accurately detected plant-pollinator, plant-herbivore, and even some mutualistic relationships, from single-interacting species to the whole interacting community. In addition, a time-series of ecological interactions can be facilitated with eDNA methods. Although PAI studies using eDNA methods remain in their infancy, to date they have identified higher numbers of taxa in several direct comparisons to DNA-based gut/bulk sampling and conventional survey methods. Therefore, research into the influencing factors of eDNA detection involved in PAI (e.g., sources and types, methodological standardization, database limitations, validation with conventional surveys, and existing ecological models) will benefit the growth of this application. Involvement of environmental RNA (eRNA) can further strengthen eDNA-based methods, provide a better understanding of complex species-interactions, and help to avoid false positive results. Thus, implementation of eDNA methods to study PAI can particularly benefit environmental biomonitoring surveys that are imperative for biodiversity health assessments.

Determining how pollinators visit plants versus how they carry and transfer pollen is an ongoing project in pollination ecology. The differences between how pollinators visit flowers versus how they carry pollen can be appreciable, but the current tools for identifying the pollens that bees carry have different strengths and weaknesses when used for ecological inference. In this study we use three methods to better understand a system of congeneric, co-flowering plants in the genus Clarkia and their bee pollinators: observations of plant-pollinator contact in the field, and two different molecular methods to estimate the relative abundance of each Clarkia pollen in samples collected from pollinators. We use these methods to investigate if observations of plant-pollinator contact in the field correspond to the pollen bees carry; if individual bees carry Clarkia pollens in predictable ways, based on previous knowledge of their foraging behaviors; and how the three approaches differ for understanding plant-pollinator interactions. We find that observations of plant-pollinator contact are generally predictive of the pollens that bees carry while foraging, and network topologies using the three different methods are statistically indistinguishable from each other. Results from molecular pollen analysis also show that while bees can carry multiple species of Clarkia at the same time, they often carry one species of pollen. Our work contributes to the growing body of literature aimed at resolving how pollinators use floral resources. We suggest our novel relative amplicon quantification method as another tool in the developing molecular ecology and pollination biology toolbox.

Invasive species are co-introduced with microbiota from their native range and also interact with microbiota found in the novel environment to which they are introduced. Host flexibility toward microbiota, or host promiscuity, is an important trait underlying terrestrial plant invasions. To test whether host promiscuity may be important in macroalgal invasions, we experimentally simulated an invasion in a common garden setting, using the widespread invasive macroalga Agarophyton vermiculophyllum as a model invasive seaweed holobiont. After disturbing the microbiota of individuals from native and non-native populations with antibiotics, we monitored the microbial succession trajectories in the presence of a new source of microbes. Microbial communities were strongly impacted by the treatment and changed compositionally and in terms of diversity but recovered functionally by the end of the experiment in most respects. Beta-diversity in disturbed holobionts strongly decreased, indicating that different populations configure more similar –or more common– microbial communities when exposed to the same conditions. This decline in beta-diversity occurred not only more rapidly, but was also more pronounced in non-native populations, while individuals from native populations retained communities more similar to those observed in the field. This study demonstrates that microbial communities of non-native A. vermiculophyllum are more flexibly adjusted to the environment and suggests that an intraspecific increase in host promiscuity has promoted the invasion process of A. vermiculophyllum. This phenomenon may be important among invasive macroalgal holobionts in general.

Over millennia, ecological and evolutionary mechanisms have shaped macroecological patterns across the tree of life. Research describing these patterns at both regional and global scales has traditionally focused on the study of metazoan species. Consequently, there is a limited understanding of cross-phylum biogeographic structuring and an escalating need to understand the macroecology of both microscopic and macroscopic organisms. Here we used environmental DNA (eDNA) metabarcoding to explore the biodiversity of marine metazoans, protists and bacteria along an extensive and highly heterogeneous coastline. Our results showed remarkably consistent biogeographic structure across the kingdoms of life despite billions of years of evolution. Analyses investigating the drivers of these patterns for each taxonomic kingdom found that environmental conditions (such as temperature) and, to a lesser extent, anthropogenic stressors (such as fishing pressure and pollution) explained some of the observed variation. Additionally, metazoans displayed biogeographic patterns that suggested regional biotic homogenization. Against the backdrop of global pervasive anthropogenic environmental change, our work highlights the importance of considering multiple domains of life to understand the maintenance and drivers of biodiversity patterns across broad taxonomic, ecological and geographical scales.

It is well established that biotic-interspecific interactions, such as competition, mutualism, parasitism or predation, modulate species dynamics in demographic and evolutionary terms. It is also acknowledged that biotic interactions can even have major effects on local population dynamics and scale-up to determine wider species' ranges (Wisz et al., 2013). Notwithstanding, the study of biotic interactions has been mostly ignored in biogeography and phylogeography research, for which it has been long assumed that abiotic factors (e.g. climate) mostly drive the ecological and evolutionary processes that underlie species' distribution. Consequently, our knowledge is scarce about the role of biotic interactions in determining spatial patterns of genetic diversity and structure. In a From the Cover article in this issue of Molecular Ecology, Ortego & Knowles (2020) address the study of positive and negative plant-plant interactions and test whether their demographic consequences translate into broadscale patterns of genomic variation in two oak species from the iconic California Floristic Province. The integrative approach undertaken in this study reveals that some models that incorporate competition or facilitation better explain genomic patterns than null models in which species respond only to variations in environmental suitability. These findings highlight the relevance of biologically informed model-based approaches for inferring the evolutionary consequences of species' range dynamics, which is of particular importance in today's global change context.

While the identification of microbial eukaryotes using molecular tools is now widespread, additional information are needed to confirm the molecular observation and make the difference between species and population variants, and therefore to better understand the biogeography of microbial eukaryotes. In this issue of Molecular Ecology, Postel et al (2020) not only used three molecular approaches to identify subgroups of Fragilariopsis kerguelensis but also morphology and physiology to better understand the relationship between the three genotypes. They revealed that (1) the three genotypes of the diatom F. kerguelensis have a negligible gene flux; and (2) two of the genotypes are geographically isolated with different physiology but still able to crossbreed; and (3) the last one is omnipresent but reproductively isolated.